Description
GenJet™ DNA In Vitro
Transfection Reagent for MCF-7 is pre-optimized for transfecting
MCF-7 cells. MCF-7 is a breast cancer cell line was isolated in 1970 from
a 69-year-old Caucasian woman. This cell line retained several
characteristics of differentiated mammary epithelium including the ability to
process estradiol via cytoplasmic estrogen receptors and the capability of
forming domes. This cell line expresses the oncogene Wnt7B. Tumor necrosis
factor alpha (TNF alpha) inhibits the growth of MCF-7 breast cancer cells.
Treatment with anti-estrogens can modulate the secretion of insulin-like growth
factor binding protein's.
Refer to the following optimal transfection conditions for maximal transfection
efficiency on MCF-7 cells. GenJet™ reagent, 1.0 ml, is sufficient for 300 to
600 transfections in 24 well plates or 50 to 100 transfections in 6 well plates.
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Summary of
Optimal Transfection Conditions:
Confluence on the
day of transfection
Cell culture
conditions
GenJet™ (µl) : DNA (µg) Ratio
Diluent for DNA and Transfection Reagent
Incubation Time to Form GenJet™/DNA
Complex
Presence of Serum/Antibiotics
during Transfection
Change Medium After Transfection
Maximal Efficiency
Transfection
Results:
Reporter Gene
Plasmid
Efficiency (GFP %) |
90%
DMEM with 4.5 g/L glucose, 10% FBS
3:1
Serum-free DMEM with 4.5 g/L glucose
15 minutes at RT
Yes
No
36 hours
EGFP
pEGFP-N3 (CMV promoter)
75% |
Storage Condition
Store at 4 °C. If stored properly, the product is stable for 12 months or longer
GenJet™ reagent is
optimized for MCF-7
cells. MCF-7 A2 cells were grown per ATCC recommended culture medium and
transfected with pEGFP-N3 by GenJet™. The efficiency was checked 36 hours post
transfection
Data Sheet
